Biology subjects

Fumagalli, L.

Publications and source records attributed to Fumagalli, L..

12 recordsLinked to original sources

bioRxiv2026

Workflow for multiplex microsatellite panel development and sample preparation for robust amplicon sequencing of low-template and degraded DNA: validation for non-invasive genotyping in three large carnivore species

High-throughput amplicon sequencing has transformed microsatellite (STR) genotyping by overcoming many of the limitations of fragment-length analysis, enabling more accurate, cost-effective, and standardized genotyping. Yet, protocols specifically designed for high-throughput sequencing (HTS)-based STR genotyping from low-template and degraded DNA remain scarce, despite the prevalence of these challenging sample types in ecological and conservation contexts. We present a methodology for the de novo development of robust STR multiplex panels together with a laboratory protocol for efficient and reliable STR genotyping by sequencing with low quantity and quality DNA samples. The protocol comprises (i) an automated bioinformatic pipeline to design large sets of short tetranucleotide markers optimized for multiplex amplicon sequencing of degraded and low-template DNA; (ii) guidelines for efficient in vitro optimization of multiplex amplification using directly low quantity/quality template DNA; and (iii) a library preparation procedure that improves detection of low-level allele signal while enabling quality assessment of STR amplicon sequencing under limiting DNA conditions. We demonstrate the approach by developing and validating STR panels for non-invasive genotyping of three large carnivore species: a 44-plex for the grey wolf (Canis lupus), a 41-plex for the Eurasian lynx (Lynx lynx), and a 30-plex for the brown bear (Ursus arctos). Multiplex performance was high, with [≥]91% of samples successfully genotyped at [≥]50% of loci (allele size range 28-110 bp across panels) and correctly assigned to known individuals, negligible levels of noise in the controls, and high discriminatory power (PIDsibs [≤]2.4 x 1e-12), also owing to sequence variation among same-length alleles at 15-50% of loci. The approach is broadly applicable to animal and plant species, a wide range of sample types, and large-scale analysis such as genetic monitoring. Our study reinforces the value of STR amplicon sequencing for ecological and conservation applications while highlighting the importance of marker design and laboratory workflows tailored to HTS-based genotyping for accurate and efficient implementation.

ecology↗

bioRxiv2026

Blockages to biodiversity data access for conservation and sustainability management

Governments, civil society organizations and businesses often lack the biodiversity data they need for decision-making and adaptive management, impacting their planning, reporting and performance. We explored the biodiversity data needs of such actors in Colombia, Ghana and Switzerland to identify factors affecting data availability and use. Responses to questionnaire surveys showed that the data types with the biggest gaps between user needs and access were progress on conservation or sustainability actions, species populations, habitat state and ecological risk. The most frequent data blockages related to inadequate resources and organizational capacity. Obstacles significantly associated with a lack of primary data included an absence of organizational biodiversity goals and monitoring systems. Problems accessing habitat quality and species abundance data were associated with data collection methods being unknown or unavailable. Businesses were more likely than other groups to need data on threats, perhaps reflecting the increasing importance of environmental risk to the corporate sector. Businesses are less likely to collect primary data or use secondary data and are significantly more likely to be unclear on what biodiversity indicators to use. Non-business organizations are significantly more likely to be unable to access data because of a lack of funding for data collection, analysis, and use. Our results highlight the need for stakeholders across sectors to work together to find common solutions to build and invest in monitoring capacity that unblocks the flow of biodiversity data.

ecology↗

bioRxiv2026

KDM: embedding DNA/RNA motifs and sequences in a shared k-mer space for unified discovery, analysis and binding prediction

Motif discovery and binding-site prediction in DNA and RNA sequences are central tasks in regulatory genomics, yet the methodological landscape is split between interpretable but rigid position weight matrices (PWMs) and high-performing but opaque machine-learning models. We present KDM, a unifying framework in which both motifs and sequences are represented as probability distributions over a shared k-mer dictionary, embedded via the Hellinger transformation. This common geometry enables motif-sequence scoring, motif-motif comparison, de novo discovery, and binding prediction with a single primitive, the Bhattacharyya coefficient. We instantiate four tools on this representation: KDMMap for positional enrichment analysis, KDMMatch for information-content-aware motif matching, KDMFind for unsupervised motif discovery via projective non-negative matrix factorization, and KDM-LRLM for binding prediction with Lasso-regularized logistic regression. Across 1,324 transcription-factor ChIP-seq and 161 RBP eCLIP experiments, KDMMap matches CentriMos motif rankings in 84% of TF and 79% of RBP experiments, and KDMMatch agrees with Tomtom on motif annotation in 74.5% of TFs. On binding prediction across four datasets covering 2,475 experiments, KDM-LRLM matches or exceeds eight deep-learning and three k-mer-based competitors. Notably, AI methods overtake k-mer methods only in the top quartile of training-set size, indicating that data scale, not architecture, drives the recent dominance of deep models. KDM provides a single interpretable representation across the full motif-analysis workflow.

bioinformatics↗

bioRxiv2026

Glacier retreat reshapes trophic networks: interaction turnover outpaces species turnover over space-time

As glaciers retreat worldwide, newly exposed terrains are rapidly colonized by plants and their associated animal communities. Although plant-animal interactions are key for biodiversity maintenance and ecosystem functioning, the ecological processes underlying the assembly and development of trophic interactions over space-time remain poorly understood. Here, we investigated the trophic niche structure of plant-arthropod interactions along a 140-year primary succession at Mont Mine glacier foreland (Switzerland). Using arthropod gut-content DNA metabarcoding, we reconstructed trophic interactions at the food web level, revealing numerous previously undetected links among 284 arthropod taxa feeding on 263 plant taxa. Trophic niche overlap among arthropods increased following glacier retreat, indicating decreasing resource partitioning and suggesting increased resource competition. Trophic niche breadth became narrower and diet species richness declined, indicating increased trophic specialization. Notably, changes in trophic interactions occurred more rapidly than shifts in species diversity or community composition of plants and arthropods. These results demonstrate that glacier retreat reorganizes trophic networks beyond simple species turnover, reshaping biotic interactions during ecosystem development. Our findings highlight trophic interactions as sensitive indicators of biodiversity change and suggest that the stability of emerging food webs may be strongly affected as glaciers vanish worldwide.

ecology↗

bioRxiv2026

Early life thermal plasticity and adaptive divergence among populations of Arctic charr (Salvelinus alpinus)

Climate warming alters the thermal environment experienced by ectotherms, whose physiological performance and fitness are constrained by temperature. Early life stages are often the temperature-sensitive phases of the life cycle, particularly in freshwater stenotherms such as the Arctic charr (Salvelinus alpinus). In this study, we used a common garden approach to compare the response and phenotypic plasticity of four charr populations to warmer conditions. These populations inhabit thermally contrasted lakes, including both native and introduced populations. We reared embryos at either an optimal (5{degrees}C) or a warmer (8.5{degrees}C) temperature. We tested adaptive divergence among populations in four traits (survival, incubation duration, body length and yolk sac volume) using Qst - Fst comparisons. We found that warmer temperature reduced body size, survival and incubation time, while thermal reaction norms differed among populations. Unexpectedly, populations originating from warmer environments did not consistently exhibit higher trait values under elevated temperatures. In contrast, the colder high-altitude population exhibited higher survival rates and lower yolk reserves for a given size under heat stress than the other populations. Our results highlight substantial variation among populations in responses to heat stress, emphasizing the complexity of predicting population responses to climate warming.

evolutionary biology↗

bioRxiv2023

Investigating fine-scale breeding habitat use by amphibians in a continuous wetland using environmental DNA

Designing effective conservation plans to protect species from extinction requires a better understanding of their ecology. Conventional methods used to investigate habitat use are time consuming, and detectability of cryptic species is often insufficient. Environmental DNA (eDNA)-based approaches now provide an alternative for ecosystems monitoring and assessment. Nevertheless, to our knowledge, such methods have never been applied to investigate habitat use at a fine scale in a continuous wetland environment. Here, we used an eDNA metabarcoding approach to characterize the breeding habitat use of local amphibian species in a wet meadow expanse along the southern shore of Lake Neuchatel, Switzerland. We retrieved DNA from six out of the seven species expected to be present. We tested the influence of six abiotic environmental variables on overall species communities as well as individual species occurrences. We showed that the main factor structuring species communities was water temperature, and that the distribution of three amphibian species was associated with several environmental variables. Our results indicate that the eDNA approach is a promising tool to study species ecology at a small scale in continuous wetland habitats.

ecology↗

bioRxiv2022

Niche overlap between two large herbivores across landscape variability using dietary eDNA metabarcoding

Understanding the relationship between a species feeding strategy and its environment (trophic ecology) is critical to assess environmental requirements and improve management policies. However, measuring trophic interactions remains challenging. Among the available methods, quantifying the plant composition of a species diet indicates how species use their environment and their associated niche overlap. Nevertheless, most studies focusing on herbivore trophic ecology ignore the influence that landscape variability may have. Here, we explored how landscape variability influences diet composition through niche overlap. We used eDNA metabarcoding to quantify the diet composition of two large herbivores of the Bialowieza Forest, Red deer (Cervus elaphus) and European bison (Bison bonasus) to investigate how increasing habitat quality (i.e. higher abundance of deciduous forage species) and predation risk (i.e. density of wolf in the area) influence their diet composition and niche partitioning. Our findings indicate diet composition is non-homogeneous across the landscape, both within and between species. Red deer showed greater diet variability and lower niche overlap within species compared to bison. We detected a reduction of niche overlap for red deer with increasing predation risk, leading to more dissimilar diets, suggesting their feeding behaviour is affected by wolf presence. This correlation was not found for bison, which are rarely predated by wolf. Higher habitat quality was associated with higher niche overlap only within bison, probably due to their suboptimal feeding strategy as browsers. These results show the importance of integrating environment-induced diet variation in studies aimed at determining the landscape usage or niche overlap of a species. Niche overlap can therefore be a powerful tool for inferring predation risk for red deer, and estimating predator abundance or its perception. This approach has enormous potential for wildlife management in areas where predator estimates are unknown or difficult to quantify.

molecular biology↗

bioRxiv2022

Hexanucleotide repeat expansions in C9orf72 alter microglial responses and prevent a coordinated glial reaction in ALS

Neuroinflammation is an important hallmark in amyotrophic lateral sclerosis (ALS). Experimental evidence has highlighted a role of microglia in the modulation of motor neuron degeneration. However, the exact contribution of microglia to both sporadic and genetic forms of ALS is still unclear. We generated single nuclei profiles of spinal cord and motor cortex from sporadic and C9orf72 ALS patients, as well as controls. We particularly focused on the transcriptomic responses of both microglia and astrocytes. We confirmed that C9orf72 is highly expressed in microglia and shows a diminished expression in carriers of the hexanucleotide repeat expansion (HRE). This resulted in an impaired response to disease, with specific deficits in phagocytic and lysosomal transcriptional pathways. Astrocytes also displayed a dysregulated response in C9orf72 ALS patients, remaining in a homeostatic state. This suggests that C9orf72 HRE alters a coordinated glial response, which ultimately would increase the risk for developing ALS. Our results indicate that C9orf72 HRE results in a selective microglial loss-of-function, likely impairing microglial-astrocyte communication and preventing a global glial response. This is relevant as it indicates that sporadic and familial forms of ALS may present a different cellular substrate, which is of great importance for patient stratification and treatment.

neuroscience↗

bioRxiv2021

Widespread Utilization of Diverse Organophosphate Pollutants by Marine Bacteria

Anthropogenic organophosphates (AOPs), such as phosphotriesters, are used extensively as plasticizers, flame retardants, nerve agents and pesticides. Soil bacteria bearing a phosphotriesterase (PTE) can degrade AOPs, but whether bacteria are capable of utilizing AOPs as a phosphorus source, and how widespread PTEs are in nature, remains unclear. Here, we report the utilization of diverse AOPs by four model marine bacteria and seventeen bacterial isolates from seawater samples. To unravel the details of AOP utilization, two novel PTEs from marine bacteria were isolated and characterized. When expressed in E. coli, these PTEs enabled growth on a pesticide analog as the sole phosphorus source. Utilization of AOPs provides bacteria with a source of phosphorus in depleted environments and offers a new prospect for the bioremediation of a pervasive class of anthropogenic pollutants. One sentence summaryWidespread utilization of diverse organophosphate pollutants by over 20 marine bacterial strains represents a new hope for ocean bioremediation.

microbiology↗

bioRxiv2020

Efficient long-range conduction in cable bacteria through nickel protein wires

Filamentous cable bacteria display unrivalled long-range electron transport, generating electrical currents over centimeter distances through a highly ordered network of fibers embedded in their cell envelope. The conductivity of these periplasmic wires is exceptionally high for a biological material, but their chemical structure and underlying electron transport mechanism remain unresolved. Here, we combine high-resolution microscopy, spectroscopy, and chemical imaging on individual cable bacterium filaments to demonstrate that the periplasmic wires consist of a conductive protein core surrounded by an insulating shell layer. The core proteins contain a sulfur-ligated nickel cofactor, and conductivity decreases when nickel is oxidized or selectively removed. The involvement of nickel as the active metal in biological conduction is remarkable, and suggests a hitherto unknown form of electron transport that enables efficient conduction in centimeter-long protein structures.

microbiology↗

bioRxiv2020

Human motor units in microfluidic devices are impaired by FUS mutations and improved by HDAC6 inhibition

Neuromuscular junctions (NMJs) ensure proper communication between motor neurons and muscle through the release of neurotransmitters. In motor neuron disorders, such as amyotrophic lateral sclerosis (ALS), NMJs degenerate resulting in muscle atrophy, paralysis and respiratory failure. The aim of this study was to establish a versatile and reproducible in vitro model of a human motor unit to study the effect of ALS-causing mutations. Therefore, we generated a co-culture of human induced pluripotent stem cell-derived motor neurons and human primary mesoangioblast-derived myotubes in microfluidic devices. A chemotactic and volumetric gradient facilitated the growth of motor neuron neurites through microgrooves resulting in the interaction with myotubes and the formation of NMJs. We observed that ALS-causing FUS mutations resulted in a reduced neurite outgrowth and in a decreased NMJ number. Interestingly, the selective HDAC6 inhibitor, Tubastatin A, improved the neurite outgrowth and the NMJ morphology of FUS-ALS co-cultures, further prompting HDAC6 inhibition as a potential therapeutic strategy for ALS.

neuroscience↗

bioRxiv2019

C9orf72-derived arginine-containing dipeptide repeats associate with axonal transport machinery and impede microtubule-based motility

Hexanucleotide repeat expansions in the C9orf72 gene are the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). How this mutation leads to these neurodegenerative diseases remains unclear. Here, we use human induced pluripotent stem cell-derived motor neurons to show that C9orf72 repeat expansions impair microtubule-based transport of mitochondria, a process critical for maintenance of neuronal function. Cargo transport defects are recapitulated by treating healthy neurons with the arginine-rich dipeptide repeat proteins (DPRs) that are produced by the hexanucleotide repeat expansions. Single-molecule imaging shows that these DPRs perturb motility of purified kinesin-1 and cytoplasmic dynein-1 motors along microtubules in vitro. Additional in vitro and in vivo data indicate that the DPRs impair transport by interacting with both microtubules and the motor complexes. We also show that kinesin-1 is enriched in DPR inclusions in patient brains and that increasing the level of this motor strongly suppresses the toxic effects of arginine-rich DPR expression in a Drosophila model. Collectively, our study implicates an inhibitory interaction of arginine-rich DPRs with the axonal transport machinery in C9orf72-associated ALS/FTD and thereby points to novel potential therapeutic strategies.

neuroscience↗